The goal of this project is to develop innovative digital microscope autofocusing techniques for automated cytogenetics applications. We propose a novel multi-resolution image analysis approach to focus measurement and detection, based on the recently developed mathematical theory of wavelet transform. In comparison to currently available single-resolution techniques, the proposed method overcomes their fundamental limitations and promises considerably more accurate, reliable and faster means to compute and determine in-focus image position for image acquisition. This will significantly increase the ability and efficacy of automated scanning microscope instruments for clinical and cancer cytogenetics applications. In Phase 1 we will investigate the feasibility of the proposed method based on its utilization in fluorescence microscopy. We will develop and implement the algorithm and software for multi-resolution focus function computation and in-focus position determination. We will test and evaluate the new method against the current best-performing algorithms by comparing (1) Accuracy; (2) Range; (3) Insensitivity to other parameters; and (4) Speed. If the new approach achieves superior performance, in Phase 2 the technique will be further developed and extended to bright-field microscopy applications. When fully developed, the new technology will be made available to Applied Imaging (AIC) for integration into the PowerGene cytogenetics automation products. PROPOSED COMMERCIAL APPLICATIONS: As soon as the new techniques are developed and qualified for routine application, they will be made available to AIC for incorporation into the PowerGene product line of cytogenetics automation equipment, both in new systems sold and as an upgrade to existing systems already in use in cytogenetics labs, thus commercializing the technology quickly.